CN113058074A - Temperature-sensitive filler composition - Google Patents
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Abstract
The invention provides a temperature-sensitive filler composition, which comprises temperature-sensitive mixed powder capable of forming temperature-sensitive gel by adding water during use, wherein the temperature-sensitive mixed powder is prepared by the following method: dispersing polymer microspheres capable of stimulating collagen generation when injected into a body in temperature-sensitive gel, wherein the temperature-sensitive gel is liquid at the ambient temperature of less than 34 ℃ and is in a gel state at the temperature of 34-37 ℃, and the dosage ratio of the polymer microspheres to the temperature-sensitive gel is 60 mg: 0.25-6 mL, preferably 60 mg: 2 mL; and freeze-drying the temperature-sensitive gel dispersed with the polymer microspheres to obtain the mixed powder. Compared with the condition of independently injecting the polymer microspheres or independently using the gel injection, the self-filling gel injection not only can immediately generate the filling effect by means of the temperature-sensitive gel, but also can continuously generate the continuous self-filling effect subsequently.
Description
Technical Field
The invention relates to a temperature-sensitive filler composition for injection for skin regeneration/filling and a preparation method and application thereof.
Background
In recent years, with the development of the cosmetic industry, cosmetic products are increasing, and cosmetic forms are upgrading, and the former is mainly surgery, and the former is mainly noninvasive filler injection. Therefore, the development of injectable filler products is continuing.
Currently, dermal fillers on the market are classified into the following three major categories depending on whether they are absorbed by the body: 1. the rapidly absorbable dermal filler mainly comprises hyaluronic acid and derivatives thereof, collagen and the like; 2. slowly absorbable dermal filler mainly comprising L-polylactic acid (PLLA) and the like; 3. the non-absorbable dermal filler mainly comprises polymethyl methacrylate (PMMA) and the like.
The collagen is a filling cosmetic material with the longest application history, mainly comes from bovine collagen, and is characterized by instant effect and rapid action. The mechanism of action is simply to increase the amount of tissue. The height of the action part is changed, thereby achieving the effects of shaping and beautifying. Unfortunately, collagen is almost completely absorbed three months after injection into the body, and its cosmetic and cosmetic effects disappear, and there is a possibility of allergic reactions. The hyaluronic acid is an injection plastic cosmetic material which is started to be applied in recent years, the plastic cosmetic action mechanism of the hyaluronic acid is similar to that of collagen, the high-quality hyaluronic acid can be stored in vivo for 3-6 months, and the hyaluronic acid is improved compared with the collagen, but the duration is still short, and the pain of the hyaluronic acid is obvious during injection. Polyacrylamide and polymethyl methacrylate are not degraded and absorbed and remain in the body for a long time, and the defects limit the use of the polyacrylamide and polymethyl methacrylate.
The levorotatory polylactic acid is recently attracting market attention as a filling material for cosmetic and cosmetic purposes. Because it is slowly degraded into non-toxic harmless carbon dioxide and water in vivo and discharged out of body, it can not cause irreversible damage to human body, so that it has good biocompatibility. Moreover, the material can stimulate the regeneration of human autologous tissues, promote the secretion of collagen and achieve the effect of autologous filling. The length of the effect is determined according to the molecular weight of the levorotatory polylactic acid, and the effect of the levorotatory polylactic acid product (Sculptra) used at present can be as long as 2-3 years. However, since the injection of L-polylactic acid does not produce an immediate cosmetic effect, it is necessary to wait until the in vivo production of self-collagen is achieved.
Chinese patent CN10425870A discloses a polylactic acid microsphere and cross-linked hyaluronic acid mixed gel for injection and a preparation method thereof. However, since the polylactic acid and the cross-linked hyaluronic acid mixed gel are in a mixed state for a long time, that is, the polylactic acid is in water for a long time, the polylactic acid is gradually hydrolyzed during the storage process, thereby affecting the therapeutic effect, and high-temperature sterilization is required to further destroy the stability and therapeutic effect of the polylactic acid microspheres during the preparation of the gel.
Disclosure of Invention
The invention aims to provide a temperature-sensitive filler composition for injection for skin regeneration/filling, a preparation method and application thereof, aiming at the defects of the prior art.
The invention provides a temperature-sensitive filler composition, which comprises a temperature-sensitive mixed powder capable of forming a temperature-sensitive gel when being added with water during use, wherein the temperature-sensitive mixed powder is prepared by the following method: dispersing polymer microspheres capable of stimulating collagen generation when injected into a body in temperature-sensitive gel, wherein the temperature-sensitive gel is liquid at the ambient temperature of less than 34 ℃ and is in a gel state at the temperature of 34-37 ℃, and the dosage ratio of the polymer microspheres to the temperature-sensitive gel is 60 mg: 0.25-6 mL, preferably 60 mg: 2 mL; and freeze-drying the temperature-sensitive gel dispersed with the polymer microspheres to obtain the mixed powder.
The lyophilization profile of freeze-drying can be as follows. Pre-freezing: cooling: the temperature of the plate layer is-45 ℃ for 15 minutes; maintaining: the temperature of the plate layer is-45 ℃ and 240 minutes. And (3) drying: vacuumizing: the temperature of a plate layer is-40 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 60 minutes; maintaining: the temperature of the plate layer is-40 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 0 minute; vacuumizing and heating: the temperature of a plate layer is-25 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 300 minutes; maintaining: the temperature of a plate layer is-25 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 1500 minutes; vacuumizing and heating: the temperature of a plate layer is-10 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is-10 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 0 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is 0 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 10 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is 10 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 28 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 15 minutes; maintaining: the temperature of the plate layer is 28 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 180 minutes.
The second aspect of the present invention also provides a temperature-sensitive filler composition, including: polymer microspheres capable of stimulating collagen production when injected into the body; the temperature-sensitive gel is liquid when the environmental temperature is lower than 34 ℃ and is in a gel state at 34-37 ℃; wherein, the polymer microsphere and the temperature sensitive gel are independently packaged and stored, and when in use, the weight ratio of the polymer microsphere to the temperature sensitive gel is 60 mg: mixing the temperature-sensitive gel in a ratio of 0.25-6 mL for injection, preferably according to a ratio of 60mg of polymer microsphere: 2mL of temperature sensitive gel was mixed for injection.
In the invention, the polymer microspheres capable of stimulating collagen generation when injected into a body and the temperature-sensitive gel composition (liquid at the temperature of lower than 34 ℃ and gel state at the temperature of 34-37 ℃) form the composite temperature-sensitive filler, so that the temperature of the human body is 36-37 ℃, the temperature-sensitive gel composition firstly directly simulates the action of collagen in the gel state to rapidly generate a filling and beautifying effect when injected into the skin, and then the dispersed polymer microspheres can continuously stimulate the organism to generate collagen to achieve an autologous filling effect. Therefore, compared with the condition of independently injecting the polymer microspheres or the condition of independently using the gel injection, the self-filling gel injection not only can immediately generate the filling effect by means of the temperature-sensitive gel, but also can continuously generate the continuous self-filling effect subsequently. In addition, in the invention, the polymer microspheres and the temperature-sensitive gel are mixed and freeze-dried to form freeze-dried powder, and water is added to form the temperature-sensitive gel for injection when the freeze-dried powder is used; or the polymer microspheres and the temperature-sensitive gel are independently packaged and stored, and are mixed for injection when in use. And the polymer microspheres are dispersed in the gel, so that the degradation speed of the polymer microspheres is delayed. Therefore, the polymer microspheres are basically in an anhydrous dry state in the preservation process before use, the shelf life of the polymer microspheres can be greatly prolonged, and when the polymer microspheres are used, the polymer microspheres are quickly dispersed in liquid temperature-sensitive gel or mixed powder containing the polymer microspheres and temperature-sensitive is dispersed in water (the liquid state is more beneficial to mixing and dispersing), and after the polymer microspheres are uniformly mixed and dispersed, the polymer microspheres can be properly heated at body temperature to form gel which can be injected into subcutaneous tissues, or injected into subcutaneous tissues in a liquid state to quickly form gel under the action of the body temperature. Thus, the polymeric microspheres have a short contact time with liquid water, reducing hydrolysis. In addition, after the composite filler is injected into a body, the polymer microspheres are dispersed in the gel to delay the degradation of the polymer to a certain extent, and the effective time of the composite filler is further prolonged.
As 60mg of polymeric microspheres: 2mL of temperature-sensitive gel is used for dosing. When the temperature-sensitive gel is used, 60mg of polymer microspheres are dispersed in 2mL of temperature-sensitive gel, on one hand, the 2mL of temperature-sensitive gel can quickly form a filling effect, on the other hand, 60mg of polymer microspheres can be uniformly dispersed in 2mL of temperature-sensitive gel composition, local concave-convex points cannot be formed, and the dispersion of the polymer microspheres in the gel can slow down the degradation of the polymer and is easy to form good connection with the filling effect of the gel. In addition, 60mg of polymer microspheres are preferably dispersed in 2mL of temperature-sensitive gel to be used as a single agent for combined packaging, so that the kit with the mass combination is convenient to finish single use.
Preferably, the polymer microspheres are at least one of polylactic acid microspheres, polydioxanone microspheres and polycaprolactone microspheres. The polylactic acid microsphere and the polycaprolactone microsphere are preferred, and the levorotatory polylactic acid microsphere with the molecular weight of 10,000-500,000 or the polycaprolactone microsphere with the molecular weight of 10,000-500,000 is more preferred. The polylactic acid microspheres and the polycaprolactone microspheres are biodegradable microspheres which can be biodegraded after being acted, and have no residue and low toxic and side effects.
Preferably, the average particle size of the polymer microspheres is 1-200 μm, preferably 20-80 μm. The polymer microspheres with the particle size range are selected, so that the polymer microspheres are good in stability, easy to rapidly and uniformly disperse in gel and free of agglomeration. The polymer microspheres have a particle size of 20 μm or more and are prevented from being phagocytized by macrophages, while the polymer microspheres have a particle size of 80 μm or less and are very easy to inject.
Preferably, the temperature-sensitive gel comprises at least one of methyl cellulose, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus, a novel high polymer material copolymerized by N-vinyl epsilon caprolactam-vinyl acetate-polyethylene glycol (57:30: 13)), poloxamer series compounds and chitosan series compounds with the mass fraction of 1-70 wt% (preferably 2-30 wt%). The temperature-sensitive gel composition comprises methylcellulose, Soluplus, poloxamer series compounds and chitosan series compounds, and can form temperature-sensitive gel by means of the characteristics of the methylcellulose, the Soluplus, the poloxamer series compounds and the chitosan series compounds. These gels are compositions that reversibly change between a liquid state and a gel state, e.g., when heated to a certain temperature, they can change from a liquid state to a gel state, and when cooled, they can change from a gel state to a liquid state. The temperature at which the liquid phase changes to the gel phase is called the gelation temperature, and the temperature at which the gel phase changes to the liquid phase is called the gel liquefaction temperature. The temperature-sensitive gel composition is liquid when the ambient temperature is lower than 34 ℃, and is in a gel state when the temperature is 34-37 ℃ (close to the body temperature of a human body).
Preferably, the temperature-sensitive gel further comprises at least one additive selected from sodium carboxymethylcellulose, sodium alginate, hyaluronic acid, amino acids, polypeptides, proteins, nucleic acids, vitamins and polysaccharides.
The product related by the invention belongs to a third class of medical appliance products, and has the following advantages: (1) compared with the pure hyaluronic acid gel, the polymer microspheres have an ultra-long biodegradation speed and long retention time in vivo, and can promote the secretion of collagen by activating fibroblasts, so that the aged skin has elasticity again, and the effect is natural, gradual, remarkable and lasting without frequent injection; (2) compared with the situation of simply using the polymer microspheres, the product is in a gel state during injection, the polymer microspheres cannot flow everywhere, the rapid filling effect is achieved, the instant short-term beautifying effect is achieved, and the polymer microspheres can be used for stimulating the production of collagen so as to achieve the purpose of lasting beautifying effect; (3) compared with polylactic acid microspheres and cross-linked hyaluronic acid mixed gel, the product is in the form of dry powder and gel solution respectively, or in the form of freeze-dried powder after the two are mixed uniformly, the polymer microspheres are basically in an anhydrous state in the preservation process before use, hydrolysis cannot be caused, and the stability of the microspheres is not influenced.
Drawings
FIG. 1 is a phase transition of mixed gel sample 1 at different temperatures, where A is room temperature and is in solution; b is a gel at 37 ℃.
FIG. 2 photomicrograph of microspheres prepared as in example 1, with the optical photomicrograph at 10 times magnification.
FIG. 3 photomicrograph of microspheres prepared as in example 2, with the optical photomicrograph at 10 times magnification.
FIG. 4 photomicrograph of microspheres prepared as in example 3, with the optical photomicrograph at 10 times magnification.
Detailed Description
The present invention will be described in detail by way of examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art may be made in light of the above teachings. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values exemplified below.
The invention relates to a temperature-sensitive filler composition for injection. In one embodiment, the temperature-sensitive filler composition mainly exists in the form of mixed freeze-dried powder, and when the temperature-sensitive filler composition is used, water is added to form temperature-sensitive composite gel for injection filling. The mixed freeze-dried powder is prepared by taking polymer microspheres and temperature-sensitive gel as raw materials, and the polymer microspheres and the temperature-sensitive gel are uniformly mixed, for example, the polymer microspheres are dispersed in the temperature-sensitive gel. In another embodiment, the polymer microspheres and the temperature-sensitive gel are separately packaged and stored, and are mixed according to a certain proportion for injection when in use.
The polymer microspheres are polymer microspheres capable of stimulating collagen production when injected into a body, such as at least one of polylactic acid microspheres, polydioxanone microspheres and polycaprolactone microspheres, and preferably levorotatory polylactic acid microspheres and polycaprolactone microspheres with excellent biocompatibility. Polylactic acid microspheres with the molecular weight of 10,000-500,000 and/or polycaprolactone microspheres with the molecular weight of 10,000-500,000 can be adopted. The polymer microspheres with the average particle size of 1-100 mu m, preferably 20-80 mu m, can be adopted. The polymer microspheres have a particle size of 20 μm or more and can be prevented from being phagocytized by macrophages, while the polymer microspheres have a particle size of 80 μm or less and can be injected very easily. And the polymer microspheres with the particle size range are selected, so that the polymer microspheres are good in stability, easy to rapidly and uniformly disperse in gel, and cannot form agglomeration.
Temperature-sensitive gels are compositions that reversibly change between a liquid state and a gel state, e.g., when heated to a certain temperature, they can change from a liquid state to a gel state, and when cooled, they can change from a gel state to a liquid state. The temperature at which the liquid phase changes to the gel phase is called the gelation temperature, and the temperature at which the gel phase changes to the liquid phase is called the gel liquefaction temperature. The temperature-sensitive gel composition is liquid when the ambient temperature is lower than 34 ℃, and is in a gel state when the temperature is 34-37 ℃ (close to the body temperature of a human body).
Temperature-sensitive gels based on methylcellulose, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymers (Soluplus), poloxamer series compounds, chitosan series compounds, and the like can be used. Methylcellulose is a nonionic cellulose ether made by the introduction of methyl groups into cellulose by etherification, and has a reverse thermal gelation property of being liquid at low temperatures and converting to a gel at elevated temperatures; soluplus, a novel high molecular material copolymerized by N-vinyl epsilon caprolactam-vinyl acetate-polyethylene glycol (57:30:13), is a copolymer with the reverse thermal gelation property of being liquid at low temperature and being changed into gel when the temperature rises; poloxamers are PEO-PPO-PEO type nonionic triblock copolymers consisting of Polyoxyethylene (PEO) and polyoxypropylene (PPO), and have the reverse thermal gelation property that the poloxamers are liquid at a low temperature and are converted into gels at a high temperature at a certain concentration; chitosan is also called chitosan and is obtained by deacetylation of chitin widely existing in the nature, and has a chemical name of polyglucosamine (1-4) -2-amino-B-D glucose. Chitosan or hydroxybutyl chitosan also has the reverse thermal gelation property of being liquid at low temperature and turning into gel at elevated temperature. Preferably, methylcellulose having a molecular weight of 18,000 to 200,000 is used. Soluplus with molecular weight of 90,0000-140,000 can be selected, and poloxamer series compounds with molecular weight of 7,680-14,600, such as poloxamer 407 and/or poloxamer 188, can be also used. And chitosan with molecular weight of 10,000-1,000,000, hydroxybutyl chitosan, carboxymethyl chitosan, etc. are used.
In the temperature-sensitive gel composition, the content of methylcellulose, Soluplus, poloxamer series compounds and/or chitosan series compounds can be 1-70 wt%, and preferably 2-30 wt%. Amounts above 2 wt% are advantageous for rapid gel formation, while amounts below 30wt% best mimic the gel state of collagen.
The temperature-sensitive gel composition is preferably formulated with water, preferably sterile water for injection. The temperature-sensitive gel composition may also include one or more additives, such as sodium carboxymethylcellulose, alginic acid, hyaluronic acid, amino acids, polypeptides, proteins, nucleic acids, vitamins, and polysaccharides.
Preferably, the temperature-sensitive gel composition with the osmotic pressure of 250-350 mOsm/L is adopted, and 0.9% sodium chloride or PBS buffer solution is adopted to adjust the osmotic pressure of the gel composition when the gel composition is prepared. The resulting gel composition is preferably subjected to sterilization/disinfection treatment, such as filtration disinfection using a 0.22 micron membrane or high temperature sterilization, preferably filtration disinfection using a 0.22 micron membrane.
The polymer microspheres can be prepared by adopting the existing method, for example, the polymer microspheres are prepared by referring to CN104258470A, the polylactic acid microspheres with the average grain diameter of 1-200 μm, preferably 20-80 μm are obtained by screening, and each ml of the polylactic acid microspheres contains 30-90 mg of polymer microspheres for packaging and standby.
The temperature-sensitive gel can also be prepared by adopting the existing gel preparation method. In the invention, methylcellulose, Soluplus, poloxamer series compounds and/or chitosan series compounds can be dispersed in water for injection, additives such as sodium carboxymethylcellulose are added according to needs, the osmotic pressure is adjusted by PBS buffer solution, and the mixture is subjected to filtration sterilization by a 0.22 micron membrane or high-temperature sterilization and then is packaged into 0.25-6 mL per unit. Preferably, the temperature sensitive gel is stored at hypothermic conditions, more preferably in liquid form at temperatures below 10 ℃.
The polymer microspheres are stored in a dry powder state, the temperature-sensitive gel is stored in a liquid state, and the two are independently packaged and stored, so that the polymer microspheres cannot be influenced by moisture or other additives in the gel during storage, and cannot be influenced by sterilization (particularly high-temperature sterilization) required by the gel. The unit polymeric microspheres and the unit gel composition can be mixed in situ at the time of use. Or dispersing the polymer microspheres in the temperature-sensitive gel, then forming freeze-dried composite powder by adopting a freeze-drying method for storage, and dispersing the composite dry powder in water to form the temperature-sensitive composite gel when in use.
When the composite gel solution is used, a composite gel solution (a composite gel solution formed by dissolving the composite dry powder in water or a composite gel solution formed by dispersing the polymer microsphere dry powder in temperature-sensitive gel) can be sucked by an injector, and the temperature-sensitive mixed gel of the polylactic acid microspheres for injection can be obtained by heating the composite gel solution by using the body temperature and can be immediately used for injection; alternatively, the polymeric microspheres and the temperature-sensitive gel can be separately placed into a pre-filled syringe having two compartments (one containing the polylactic acid microspheres and the other containing the temperature-sensitive gel). Before injection, the gel solution passes through the intermediate diaphragm to be fully mixed with the polylactic acid microspheres, and the temperature of the mixture is raised by using the body temperature to obtain the temperature-sensitive composite gel of the polylactic acid microspheres for injection.
The ratio of polymer microspheres can be as 60 mg: and (3) compounding 0.25-6 mL of temperature-sensitive gel. When the temperature-sensitive gel is used, 60mg of polymer microspheres are dispersed in 2mL of temperature-sensitive gel, on one hand, the 2mL of temperature-sensitive gel can quickly form a filling effect, on the other hand, 60mg of polymer microspheres can be uniformly dispersed in 2mL of temperature-sensitive gel composition, local concave-convex points cannot be formed, and the dispersion of the polymer microspheres in the gel can slow down the degradation of the polymer and is easy to form good connection with the filling effect of the gel. In addition, 60mg of polymer microspheres are preferably dispersed in 2mL of temperature-sensitive gel to be used as a single agent for combined packaging, so that the kit with the mass combination is convenient to finish single use. When the composite dry powder (polylactic acid microsphere mixed temperature-sensitive gel dry powder) is adopted, the use amount of the composite dry powder is that the amount ratio of the added water to the polymer microsphere can be referred to, namely, 60mg of the mixed dry powder is added into 2mL of water. It should be understood that the water, including the water with the temperature-sensitive gel, is preferably water that has been subjected to a sterilization treatment.
The present invention is further illustrated by the following examples, which do not limit the scope of the present invention. The lyophilization profile of the freeze-drying used in the examples can be carried out by the following process, which is not repeated in the examples. Pre-freezing: cooling: the temperature of the plate layer is-45 ℃ for 15 minutes; maintaining: for 240 minutes. And (3) drying: vacuumizing: the temperature of a plate layer is-40 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 60 minutes; maintaining: the temperature of the plate layer is-40 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 0 minute; vacuumizing and heating: the temperature of a plate layer is-25 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 300 minutes; maintaining: the temperature of a plate layer is-25 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 1500 minutes; vacuumizing and heating: the temperature of a plate layer is-10 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is-10 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 0 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is 0 ℃, the vacuum degree is less than or equal to 0.14mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 10 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 30 minutes; maintaining: the temperature of a plate layer is 10 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 30 minutes; vacuumizing and heating: the temperature of a plate layer is 28 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 15 minutes; maintaining: the temperature of the plate layer is 28 ℃, the vacuum degree is less than or equal to 0.007mbar, and the time is 180 minutes.
Example 1
Adding 2g of L-polylactic acid (PLLA) with the molecular weight of 500 and 00 into 16mL of dichloromethane for dissolving, then dropwise adding into 400mL of a polyvinyl alcohol (the viscosity is 22.6mPa.s) aqueous solution with the mass fraction of 1%, stirring at normal temperature for 1 hour after dropwise adding, heating to 45 ℃, removing dichloromethane to obtain a reaction solution, filtering the reaction solution to collect a filter cake, screening out microspheres with the average particle size of 20-80 mu m, performing vacuum drying to obtain 1.3g of polylactic acid microspheres with the average particle size of 20-80 mu m, and subpackaging the microspheres into 60mg of microsphere sample 1 per bottle for later use. See fig. 2 for an optical micrograph thereof.
Placing 2g of methylcellulose (SM 400) and 0.5g of sodium alginate in a 200mL beaker, adding 60mL of hot water at 90 ℃, stirring for dissolving, and storing in a refrigerator at 4 ℃ for later use; adding 20mL (0.4g/mL) of polyethylene glycol and 14mL (0.25g/mL) of sodium citrate into the solution under stirring, adjusting the pH to 7.4, fixing the volume to 100mL, stirring at 4 ℃ for 12 hours, heating and sterilizing the temperature-sensitive gel, subpackaging into 2mL of temperature-sensitive gel samples 1 in each bottle, and standing at 4 ℃ for later use.
And (3) placing the subpackaged methylcellulose temperature-sensitive gel in a refrigerator at 4 ℃ for 1 hour, taking 2mL of the gel, injecting the gel into the subpackaged 60mg of L-polylactic acid microspheres, and shaking up to obtain the methylcellulose temperature-sensitive mixed gel of the L-polylactic acid microspheres for injection.
Example 2
Adding 2g of racemic polylactic acid (PDLLA) with the molecular weight of 15000 into 4mL of dichloromethane for dissolving, then dropwise adding into 200mL of a polyvinyl alcohol (viscosity is 22.6mPa.s) aqueous solution with the mass fraction of 1%, stirring at normal temperature for 1 hour after dropwise adding, heating to 45 ℃, removing dichloromethane to obtain a reaction solution, filtering the reaction solution to collect a filter cake, screening out microspheres with the average particle size of 20-80 mu m, and performing vacuum drying to obtain 1.4g of polylactic acid microspheres with the average particle size of 20-80 mu m as a microsphere sample 2 for later use; or split into 60mg microspheres per vial sample 2 for use. See fig. 3 for an optical micrograph thereof.
Placing 20g of Soluplus in a 100mL beaker, adding 80mL of 0.05M PBS (phosphate buffer solution) with pH7.4, stirring for dissolving, fixing the volume to obtain 100mL of gel, heating and sterilizing the temperature-sensitive gel to obtain a temperature-sensitive gel sample 2, and placing the temperature-sensitive gel sample at 4 ℃ for storage for later use; or subpackaging into 2mL of temperature-sensitive gel sample per bottle, and storing at 4 ℃ for later use.
Putting the polylactic acid microspheres prepared by the method into Soluplus temperature-sensitive gel, uniformly stirring (the dosage ratio of the polylactic acid microspheres to the Soluplus temperature-sensitive gel is 60 mg: 2mL), subpackaging into 2mL temperature-sensitive composite gel samples per bottle, and then freeze-drying to obtain polylactic acid microsphere mixed temperature-sensitive gel dry powder. Before use, injecting 2mL of injection water into the polylactic acid microsphere mixed temperature-sensitive gel dry powder, shaking and shaking uniformly to obtain the polylactic acid microsphere mixed temperature-sensitive gel.
Or placing the split Soluplus temperature-sensitive gel in a refrigerator at 4 ℃ for 1 hour, taking 2mL of the gel, injecting the gel into the split 60mg of polylactic acid microspheres, shaking up by shaking, and obtaining the Soluplus temperature-sensitive mixed gel of the polylactic acid microspheres for injection.
Example 3
Adding 2g of Polycaprolactone (PCL) with the molecular weight of 50,000 into 12mL of dichloromethane for dissolving, then dropwise adding into 200mL of a polyvinyl alcohol (the viscosity is 22.6mPa.s) aqueous solution with the mass fraction of 1%, stirring at normal temperature for 1 hour after dropwise adding, heating to 45 ℃, removing dichloromethane to obtain a reaction solution, filtering the reaction solution to collect a filter cake, screening out microspheres with the average particle size of 20-80 mu m, and performing vacuum drying to obtain 1.5g of polycaprolactone microspheres with the average particle size of 20-80 mu m as a microsphere sample 3 for later use; or split into 60mg microspheres per bottle of sample 3 for use. See fig. 4 for an optical micrograph thereof.
Putting 18g of poloxamer 407, 2g of poloxamer 188, 0.5g of sodium carboxymethylcellulose (viscosity is 12000mPa.s) and 0.25g of polyethylene oxide (PEO) with average molecular weight of 500 ten thousand into a 100mL beaker, adding 80mL of 0.9% sodium chloride, stirring for dissolving, fixing the volume to obtain 100mL of gel, heating and sterilizing the temperature-sensitive gel to obtain a temperature-sensitive gel sample 3, and storing at 4 ℃ for later use; or subpackaging into 2mL of temperature-sensitive gel sample 3 per bottle, and storing at 4 ℃ for later use.
Putting the polycaprolactone microspheres prepared by the method into poloxamer 407 temperature-sensitive gel, uniformly stirring (the dosage ratio of the polycaprolactone microspheres to the poloxamer 407 temperature-sensitive gel is 60 mg: 2mL), subpackaging into 2mL temperature-sensitive composite gel samples per bottle, and freeze-drying to obtain polycaprolactone microsphere mixed temperature-sensitive gel dry powder. Before use, the polycaprolactone microsphere mixed temperature-sensitive gel dry powder is injected into 2mL of injection water, and the polycaprolactone microsphere mixed temperature-sensitive gel is obtained by shaking up.
Or placing the packaged poloxamer 407 temperature-sensitive gel in a refrigerator at 4 ℃ for 1 hour, taking 2mL of the gel, injecting the gel into the packaged 60mg polycaprolactone microspheres, and shaking up by shaking to obtain the poloxamer 407 temperature-sensitive mixed gel of the polycaprolactone microspheres for injection.
Taking the prepared polylactic acid microsphere methyl cellulose temperature-sensitive mixed gel as a mixed gel sample 1; taking the prepared polylactic acid microsphere Soluplus temperature-sensitive mixed gel as a mixed gel sample 2; the poloxamer 407 temperature-sensitive mixed gel of the polycaprolactone microspheres prepared was used as mixed gel sample 3. And (3) adding 2mL of injection water instead of the temperature-sensitive gel into the polylactic acid microspheres, and shaking up to obtain a reference sample.
Determination of gelling temperature and time of mixed gel sample: placing the mixed gel samples 1, 2 and 3 and the reference sample in a constant-temperature water bath respectively, heating up the mixed gel samples successively, wherein the temperature is raised by 1 ℃ each time, the mixed gel samples reach the specified temperature each time and are stabilized for 10 minutes, judging according to the principle of flow (sol) -no-flow (gel), recording the temperature and the gelling time, and the results are shown in table 1.
TABLE 1 gelling temperature and time for the samples
| Sample number | Gel forming temperature (. degree. C.) | Gel forming time (minutes) |
| Mixed gel sample 1 | 37 | 8 |
| Mixed gel sample 2 | 37 | 5 |
| Mixed gel sample 3 | 34 | 3 |
| Reference sample | Does not form glue | Does not form glue |
Claims (9)
1. The temperature-sensitive filler composition is characterized by comprising temperature-sensitive mixed powder which can form temperature-sensitive gel when being added with water during use, and the temperature-sensitive mixed powder is prepared by the following method:
dispersing polymer microspheres capable of stimulating collagen generation when injected into a body in temperature-sensitive gel, wherein the temperature-sensitive gel is liquid at the ambient temperature of less than 34 ℃ and is in a gel state at the temperature of 34-37 ℃, and the dosage ratio of the polymer microspheres to the temperature-sensitive gel is 60 mg: 0.25-6 mL, preferably 60 mg: 2 mL; and
and freeze-drying the temperature-sensitive gel dispersed with the polymer microspheres to obtain the mixed powder.
2. A temperature-sensitive filler composition, characterized in that the temperature-sensitive filler composition comprises:
polymer microspheres capable of stimulating collagen production when injected into the body; and
the temperature-sensitive gel is liquid when the environmental temperature is lower than 34 ℃ and is in a gel state at 34-37 ℃;
wherein, the polymer microsphere and the temperature sensitive gel are independently packaged and stored, and when in use, the weight ratio of the polymer microsphere to the temperature sensitive gel is 60 mg: mixing the temperature-sensitive gel in a ratio of 0.25-6 mL for injection.
3. The temperature-sensitive filler composition according to claim 1 or 2, wherein the polymer microspheres are at least one of polylactic acid microspheres, polydioxanone microspheres, and polycaprolactone microspheres.
4. The temperature-sensitive filler composition according to claim 3, wherein the polymer microspheres are L-PLA microspheres with a molecular weight of 10,000-500,000 or polycaprolactone microspheres with a molecular weight of 10,000-500,000.
5. The temperature-sensitive filler composition according to any one of claims 1 to 4, wherein the polymer microspheres have an average particle diameter of 1 to 200 μm.
6. The temperature-sensitive filler composition according to claim 5, wherein the polymer microspheres have an average particle size of 20 to 80 μm.
7. The temperature-sensitive filler composition according to any one of claims 1 to 6, wherein the temperature-sensitive gel comprises at least one of methylcellulose, a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, a poloxamer series compound, and a chitosan series compound in a mass fraction of 1 to 70 wt%.
8. The temperature-sensitive filler composition according to claim 7, wherein the temperature-sensitive gel comprises at least one of methylcellulose, a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, a poloxamer series compound and a chitosan series compound in a mass fraction of 2 to 30 wt%.
9. The temperature-sensitive filler composition according to any one of claims 1 to 8, wherein the temperature-sensitive gel further comprises at least one additive selected from the group consisting of sodium carboxymethylcellulose, alginic acid, hyaluronic acid, amino acids, polypeptides, proteins, nucleic acids, vitamins and polysaccharides.
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| CN116712609A (en) * | 2023-07-26 | 2023-09-08 | 鑫华微(厦门)生物科技有限公司 | Injectable temperature-sensitive composite hydrogel and application thereof in preparation of molding and/or repairing filling material |
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